TSIV and other iridovirus homolog analysis

# Load libraries
library(tidyverse); theme_set(theme_bw())

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library(fs)

# Input files
acc_file <- "/labs/kingsley/ambenj/TSIV/resources/240814_genomes_for_core_gene_analysis.txt"
core_genes_file <- "/labs/kingsley/ambenj/TSIV/resources/Eaton2007_iridovirus_core_genes.txt"
blastp_folder <- "/labs/kingsley/ambenj/TSIV/analysis/assembly/04_blastp_iridoviruses"

# Output files
core_gene_outfile <- "/labs/kingsley/ambenj/TSIV/analysis/assembly/04_blastp_iridoviruses/core_genes_needs_curation.txt"
genbank_gene_outfile <- "/labs/kingsley/ambenj/TSIV/analysis/assembly/04_blastp_iridoviruses/core_genes_genbank.txt"
TSIV_gene_outfile <- "/labs/kingsley/ambenj/TSIV/analysis/assembly/04_blastp_iridoviruses/core_genes_TSIV.txt"

# Read in accession file
acc <- read_tsv(acc_file) %>% 
  mutate(virus_accession = str_remove(File_prefix, '\\.1'))

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acc

# Read in core genes file
core_genes <- read_tsv(core_genes_file)

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core_genes

Get Eaton sequences for manual curations

# Read blastp results files
files <- dir_ls(path = blastp_folder, glob = "*_blastp.tsv")

# function to add file name to dataframe
read_and_record_filename <- function(filename){
  read_tsv(filename, col_names = c("qseqid", "sseqid", "evalue", "stitle", "pident", "length", "mismatch", "gapopen", "qstart", "qend", "sstart", "send", "bitscore")) %>%
  mutate(filename = path_file(filename))
  }

# gather files into one dataframe
blastp <- map_df(files, read_and_record_filename) %>% 
  separate(qseqid, into=c("TSIV_ORF"), sep="\\|", remove=FALSE, extra="drop") %>% 
  mutate(TSIV_ORF = as.integer(str_remove(TSIV_ORF, "gene_"))) %>% 
  separate(filename, into=c("virus_accession"), sep="\\.", extra="drop")

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blastp %>% 
  arrange(TSIV_ORF)

# Get top hit for each accession for each TSIV ORF
top_blastp <- blastp %>%
  filter(evalue < 0.1) %>% 
  group_by(TSIV_ORF, virus_accession) %>% 
  arrange(evalue) %>% 
  slice(1) %>% 
  ungroup() %>% 
  arrange(TSIV_ORF)
top_blastp

# Get core gene table
core_genes_blastp <- top_blastp %>% 
  select(qseqid, TSIV_ORF, sseqid, virus_accession) %>% 
  pivot_wider(id_cols = c("qseqid", "TSIV_ORF"), names_from = virus_accession, values_from = sseqid) %>% 
  separate(FV3_AY548484, into=c("FV3"), sep="\\(", remove=FALSE, extra="drop") %>% 
  left_join(core_genes, ., by=c("FV3")) %>% 
  filter(!is.na(Order), !TSIV_ORF %in% c(82, 29)) %>% 
  arrange(Order)

core_genes_blastp

core_genes_blastp %>% 
  write_tsv(core_gene_outfile)

# Get TSIV core genes
TSIV_core_genes_blastp <- core_genes_blastp%>% 
  select(Order, `Gene Name`, TSIV_ORF, qseqid)
TSIV_core_genes_blastp

Get genbank sourced annotations and TSIV for core genes

# Get top results for Genbank annotation viruses
genbank_core_genes <- top_blastp %>% 
  left_join(., acc, by="virus_accession") %>% 
  filter(Annotations=="Genbank", Core_protein_alignment=="yes") %>% 
  left_join(TSIV_core_genes_blastp, .)

## Joining with `by = join_by(TSIV_ORF, qseqid)`

genbank_core_genes

# Check all expected sequences are present
genbank_core_genes %>% 
  select(Order, `Gene Name`, qseqid, TSIV_ORF, sseqid, virus_accession) %>% 
  pivot_wider(id_cols = c("Order", "Gene Name", "qseqid", "TSIV_ORF"), names_from = virus_accession, values_from = sseqid)

# Write output in long format to get protein sequences for genbank accessions
genbank_core_genes %>% 
  select(Order, `Gene Name`, virus_accession, sseqid) %>% 
  arrange(virus_accession) %>% 
  write_tsv(genbank_gene_outfile)

 # Write output in long format to get protein sequences for TSIV
TSIV_core_genes_blastp %>% 
  select(Order, `Gene Name`, qseqid) %>% 
  write_tsv(TSIV_gene_outfile)